331 EFFECT OF SUCESSIVE LAPAROSCOPIC OOCYTE RECOVERY AFTER HORMONAL TREATMENT IN CROSSBRED GOATS

2010 ◽  
Vol 22 (1) ◽  
pp. 321
Author(s):  
S. R. G. Avelar ◽  
K. C. Almeida ◽  
A. F. Pereira ◽  
F. C. Sousa ◽  
R. R. Moura ◽  
...  
Keyword(s):  
Ovarian Stimulation ◽  
Prostaglandin F2α ◽  
Hormonal Treatment ◽  
Ovarian Response ◽  
Cumulus Cells ◽  
Embryo Production ◽  
Hormonal Stimulation ◽  
Exact Test ◽  
Oocyte Recovery

Laparoscopic oocyte recovery (LOR) is a valuable tool for obtaining oocytes for in vitro embryo production. When preceded by a treatment of ovarian stimulation, this technique produces an increase in the amount of oocytes recovered. However, a little information has been found to respect the effect of successive hormonal treatments on both oocyte quantity and quality. Therefore, the objectives of this study were to evaluate the ovarian response and quantitative and qualitative COC production. Five adult crossbred goats were hormonally treated with intravaginal sponges containing 60 mg of medroxyprogesterone acetate (MAP, Progespon, Syntex, Buenos Aires, Argentina) for 11 days. In the 8th day of progestagen treatment, 50 μg of prostaglandin F2α analogue (Ciosin, Coopers, São Paulo, Brazil) was administered by i.m. injection. At this time, ovarian stimulation was initiated by the administration of 120 mg pFSH (Folltropin-V, Vetrepharm, Canada) distributed in five decreasing doses (30/30, 20/20, 20 mg), at 12-h intervals. The animals were fasted for 24 h before the laparoscopic procedure, which was performed just after the sponge removal. A laparoscopic system connected to a 22-gauge needle (WTA, Watanabe, Brazil) and a vacuum pump (Biovacuum, Biocom, Brazil) providing 30 mm Hg was used. All follicles with a size larger than 2 mm present in both ovaries were counted and aspirated. The collection medium was TCM-199 supplemented with HEPES (10 mM), heparin (20 IU mL-1), and gentamicin (40 μg mL-1). The COCs were graded based on presence of cumulus cells and cytoplasm homogeneity (I to IV). The hormonal treatment and LOR procedures were repeated three times at 14-day intervals. Data were expressed in percentage or mean ± SEM. The differences were analyzed using ANOVA and Tukey’s or Fischer’s exact test when appropriate, with P < 0.05. No statistical differences were found (P > 0.05) for the number of follicles obtained in each LOR session: 17.0 ± 3.91, 18.75 ± 2.59, and 18.0 ± 4.73, respectively. Repeated LOR procedures also did not affect (P > 0.05) the number of aspirated follicle (15.0 ± 3.92, 15.5 ± 2.33, and 16.0 ± 4.36), resulting from the three sessions, respectively. Average recovery rates were not statistically different (P > 0.05), resulting in 74.7%, 62.9%, and 64.6% between sessions. With respect to the percentage of viable COCs (GI and GII) were not observed statistical differences (P > 0.05), as verified the follow values at 1st to 3rd sessions: 76.79%, 84.62%, and 74.19%. In conclusion, three successive hormonal stimulation LOR procedures did not cause detrimental effects on quantitative and qualitative oocyte production, suggesting that this protocol can be used for programs of in vitro goat embryo production. This study was supported the following Brazilian agencies: FINEP, CNPq, FUNCAP, and CAPES.

316 EVALUATION OF A HORMONAL PROTOCOL FOR LAPAROSCOPIC OVUM PICK-UP IN PREPUBERTAL EWES

2013 ◽  
Vol 25 (1) ◽  
pp. 305
Author(s):  
P. P. M. Teixeira ◽  
L. C. Padilha ◽  
A. S. L. da Silva ◽  
F. F. P. C. Barros ◽  
L. N. Coutinho ◽  
...  
Keyword(s):  
Ovarian Stimulation ◽  
Ovulation Induction ◽  
High Performance ◽  
In Vitro Maturation ◽  
Prostaglandin F2α ◽  
Hormonal Treatment ◽  
Ovarian Response ◽  
The Other ◽  
Genetic Value

In vitro embryo production is a cutting-edge technology in constant evolution and has been used routinely. With the objective of improving genetic value, recent studies have been focused on the production of high-performance descendants from early prepubertal animals. The aim of this study was to compare hormonal protocols of ovarian stimulation for laparoscopic ovum pick-up in prepubertal sheep, to determine ovarian response and number of oocytes. For this study, 36 Santa Ines sheep, aged between 4 and 8 weeks, were submitted to a progestogen-based, short-term ovulation induction protocol (without use of prostaglandin F2α), and following 36 h, ovarian stimulation (300 IU of eCG) associated with different regimens of administration of FSH, according to experimental group. The ewes were divided into 1 of 6 groups: 2 groups received 80 mg of FSH by either single administration in the first group (G80U) or multiple constant administrations at 12-h intervals in the second group (G80M); animals belonging to 2 other groups received 160 mg in the same fashion, in a single-dose regimen (group G160U) or in multiple administrations (group G160M). The other 2 groups constituted the control groups: one received no hormonal treatment for ovulation induction and ovarian stimulation (GCN); the other received the ovulation induction protocol, but no ovulation stimulation was carried out (GCI). The animals were submitted to laparoscopic ovum pick-up 48 h following the beginning of the ovarian stimulation. The number of follicles viewed (FV), aspirated (FA), and recovered (OR) laparoscopically was recorded. Data were assessed using the one-way ANOVA test, and the comparison among groups was carried out using the Tukey test. The mean of follicles visualised, aspirated, and oocytes recovered are shown in Table 1. Group G160U had a better result regarding OR compared with the other groups. Moreover, the current study highlights that laparoscopic ovum pick-up for in vitro maturation using prepubertal ewes is viable for commercial purposes. More studies are required in order to improve the quality of oocytes and successful in vitro maturation rates. Table 1.Results of visualised follicles (VF), aspirated follicles (AF), and oocytes retrieved (OR)

169 THE RELATIONSHIP BETWEEN OOCYTE RECOVERY AND EMBRYO PRODUCTION IN BOS INDICUS

2012 ◽  
Vol 24 (1) ◽  
pp. 196
Author(s):  
M. P. Palhão ◽  
E. R. Oliveira ◽  
M. M. Gioso ◽  
B. C. Carvalho ◽  
L. G. B. Siqueira ◽  
...  
Keyword(s):  
Body Condition Score ◽  
Bos Indicus ◽  
Pregnancy Rates ◽  
Embryo Production ◽  
Exact Test ◽  
Oocyte Recovery ◽  
Conversion Rates ◽  
Condition Score ◽  
The Relationship

The ovarian follicular population has been used as a parameter to evaluate fertility and also the potential of donors undergoing assisted reproductive procedures in both human medicine and animal practice. There is a high correlation between follicular population and oocyte recovery by ovum pickup (OPU), but the relationship between oocyte recovery, embryo production and pregnancy rates may not be fully understood. The aim of the present study was to evaluate whether the conversion rate of oocytes to embryos and further pregnancies could be positively related to the number of cumulus–oocyte complexes (COC) recovered after OPU in cattle. For this purpose, records of 626 OPU sections from 251 nonlactating Gyr cows (dairy Zebu breed) were analysed. The animals had a good body condition score, were kept in a good feeding pasture (Brachiaria spp.) and were supplemented with corn silage and a mixture of corn, soybeans and vitamin and minerals, according to their nutritional requirements. For each ovarian aspiration, the ovarian follicular wave was previously synchronized with an auricular implant (Norgestomet-Crestar®), IM injections of 2 mg of oestradiol benzoate (Gonadiol®) and 0.25 mg of D-cloprostenol (Sincrocio®). The OPU procedures were performed using an ultrasound device (Aquila Vet, Esaote, São Paulo, Brazil) equipped with a vaginal sector 7.5-MHz probe, disposable 20 G needles and a vacuum pressure of 80 mmHg. The cows were ranked in quartiles regarding the total number of COC recovered. To reduce bias related to the eventual fluctuation of OPU results, for the present analysis the authors used only the recorded OPU session of each cow with the highest number of COC recovered. Viable COC were fertilized with sex-sorted (X) semen of Gyr bulls previously tested for in vitro embryo production. Conversion rates (%) of the total and viable oocytes to embryos, viable oocytes to pregnancy and embryo to pregnancy were evaluated for each quartile. Differences between the first and fourth quartiles were accessed by Fisher's exact test. In the 251 OPU, 4246 total and 3173 viable COC were recovered, resulting in the production of 1001 embryos (31.5%) and 453 pregnancies (45.3%). The cows ranked in the first, second, third and fourth quartiles produced >30 (41.6 ± 10.6), 21 to 30 (25.2 ± 3.0), 12 to 20 (15.9 ± 2.6) and <12 (6.7 ± 3.1) total oocytes. The average viable oocyte (29.1 ± 11.0, 18.1 ± 5.3, 11.1 ± 3.7 and 4.5 ± 2.7, respectively) and embryo production (8.6 ± 5.7, 5.2 ± 3.6, 3.8 ± 2.8 and 1.8 ± 1.8, respectively) were different (P < 0.0001) among all quartiles. Pregnancy rates, however, did not differ (46.0, 44.9, 43.9 and 45.6%, respectively; P > 0.05). Interestingly, the conversion rates (viable oocytes to embryos and viable oocytes to pregnancies) were higher (P < 0.0001 and 0.002) in cows from the last quartile (51.1 and 31.9%) compared with those from the first quartile (23.7 and 14.7%). In conclusion, the number of COC recovered by OPU (and consequently the ovarian follicular count) can further predict the total number of embryos and pregnancies produced, but it is not directly related to the oocyte development potential. Biotran and Fapemig Project CVZ APQ 01654/09 and BPD 0007/10.

328 IN VITRO MATURATION OF OOCYTES FROM Canindé GOATS SUBMITTED TO DIFFERENT HORMONAL TREATMENTS FOR OVARIAN STIMULATION

2010 ◽  
Vol 22 (1) ◽  
pp. 320
Author(s):  
K. C. Almeida ◽  
A. F. Pereira ◽  
A. S. Alcântara Neto ◽  
S. R. G. Avelar ◽  
F. C. Sousa ◽  
...  
Keyword(s):  
Ovarian Stimulation ◽  
Polar Body ◽  
Hormonal Treatment ◽  
Oocyte Quality ◽  
Exact Test ◽  
Gentamicin Sulfate ◽  
First Polar Body ◽  
The One ◽  
Hormonal Treatments

Oocyte IVM is a long process during which oocytes acquire their ability to support the stages of development in a stepwise manner, ultimately reaching activation of the embryonic genome. The overall success of this process can be affected by factors such as hormonal treatment for ovarian stimulation. Thus, the current study aims to evaluate the possible effects of the ovarian stimulatory protocols on the goat oocyte quality and IVM rate. Adult and cyclic Canindé goats were heat-synchronized by means of intravaginal sponges impregnated with 60 mg medroxyprogesterone acetate (MAP, Progespon, Syntex, Buenos Aires, Argentina) inserted for 11 days coupled with a luteolytic injection of 50 μg cloprostenol (Ciosin, Coopers, São Paulo, Brazil) in the 8th day of treatment. The ovarian stimulation was carried out using one of the following protocols: a) standard multi-doses (MD) with 120 mg pFSH (Folltropin-V, Vetrepharm, Canada) distributed in five injections (30/30; 20/20; 20 mg) at 12 h intervals (n = 18); b) three- doses (TD) with 120 mg pFSH administered in three injections (60; 40; 20 mg) at 24 h intervals (n = 17); c) one shot (OD) of 70 mg pFSH plus 200 IU of eCG (Novormon, Syntex) administered 36 h before sponge removal (n = 17). In MD andTD groups, the pFSH injections started in Day 8 of progestagen treatment. The follicles were aspirated just after the sponge removal using laparoscopic oocyte recovery (LOR). This procedure was performed with a 22-gauge needle and a vacuum pump at 30 mmHg. The collection medium was TCM-199 supplemented with HEPES (10 mM), heparin (20 IU mL-1), and gentamicin sulfate (40 μg mL-1). COCs were classified as grade I, II, III, or IV based on visual criteria (Baldassarre H et al. 2003 Theriogenology 56, 831-839). Good quality oocytes (grade I and II) were incubated in TCM-199 supplemented with cysteamine (100 μM), EGF (10 ng mL-1) and gentamicin sulfate (40 μgm L-1) at 38.5°C in a humidified atmosphere with 5% CO2 in air for 24 h. Oocyte maturation was assessed by the visualization of first polar body under inverted microscope. Data were expressed as percentages and analyzed using the Fischer’s exact test. No statistical differences among hormonal treatments (P > 0.05) were observed for the percentage of the good quality oocytes, with 70.4 ± 3.0% of COCs graded in I and II. The IVM rate inTD (31.4%) was statistically lower than MD (31.4% v. 46.5%, P = 0.04) group. However, no significant differences (P = 0.89) were observed between OD (45.2%) and MD groups. Thus, current results indicate that oocyte production for IVM can be facilitated using ovarian stimulation with the one shot FSH/eCG regime without affecting meiotic competence. In summary, OD and MD treatments can be used for oocyte IVM in an embryo production programme in Canindé goats. This study was supported by the following Brazilian agencies: FINEP, CNPq, FUNCAP, and CAPES.

scholarly journals Pilot study: Alternative spliced variants of the Follicle-Stimulating Hormone Receptor gene in Greek women undergoing In Vitro Fertilisation (IVF)

2019 ◽  
Vol 18 (4) ◽  
pp. 155-162
Author(s):  
Eudoxia Mamas ◽  
Despoina Mavrogianni ◽  
Rami Raouasnte ◽  
Spyros Karkatzoulis ◽  
Emmanouela Liokari ◽  
...  
Keyword(s):  
Ovarian Stimulation ◽  
Hormone Receptor ◽  
Splice Variants ◽  
Receptor Gene ◽  
Ovarian Response ◽  
Cumulus Cells ◽  
Response To Treatment ◽  
In Vitro Fertilisation ◽  
Nucleotide Polymorphisms

Assisted Reproduction Technology (ART) has proven to be a valuable tool for infertile couples. Unfortunately, no perfect ovarian stimulation protocol has been designed and ovarian response shows great variability among women. Pharmacogenomics aims at detecting genetic markers so as to individualize protocols in order to maximize ovarian response to treatment. Follicle Stimulation hormone receptor (FSHR) mutations and single nucleotide polymorphisms have been extensively studied. Splice variants of the FSHR have been detected in women undergoing in vitro fertilisation (IVF). This study aims to determine the presence FSH splice variants in Greek women undergoing IVF. RNA was extracted from cumulus cells from 35 women and analysed by real time PCR. Splice variants were detected by gel electrophoresis. Three cases of deletion of exon 9 and 2 cases of insertion of intron 8 were detected in our study group. No association between the presence of splice variants and response to ovarian stimulation was detected. Two live births were detected one in each variant group. Even though these two types of splice variant detected do not show any clinical correlation it is believed that variants of the FSHR may be associated with poor or high response to exogenous gonadotrophin so further research is necessary.

141 SUPERIORITY OF FEMALE EMBRYO PRODUCTION SYSTEM BY IN VIVO-MATURED OOCYTE AND X-SORTED SPERM IN BROWN SWISS COWS

2014 ◽  
Vol 26 (1) ◽  
pp. 184
Author(s):  
T. Yamanouchi ◽  
H. Matsuda ◽  
M. Ohtake ◽  
K. Masaki ◽  
E. Horiguchi ◽  
...  
Keyword(s):  
Prostaglandin F2α ◽  
Cumulus Cells ◽  
Embryo Production ◽  
Female Embryo ◽  
Brown Swiss ◽  
Embryo Recovery ◽  
Matured Oocyte ◽  
Matured Groups

Embryo transfer using a female embryo is an effective tool for offspring production on dairy industry; however, embryo production by embryo recovery (ER) using X-sorted semen is not sufficient because non-fertilized oocytes are recovered frequently. In Holstein cows, we developed a system for high blastocyst production that was performed by IVF using X-sorted sperm and in vivo-matured oocytes obtained by ovum pickup (OPU) after superstimulation. The purpose of this study was to adjust this system to Brown Swiss cows, comparing between ER and embryo production from oocytes derived from OPU with or without superstimulation. In the ER group, cows (n = 10) received a CIDR (Day 0) and 2 mg of oestradiol-benzoate on Day 1. A total of 30 Armour Units of FSH were injected into cows twice a day, with decreasing doses from the evening of Day 5 to the morning of Day 9. On the evening of Day 7 or 8, 0.75 mg of prostaglandin F2α (cloprostenol) was injected. The CIDR was removed on Day 8 or 9 and 0.2 mg of gonadotropin-releasing hormone (GnRH; fertirelin acetate) were injected on Day 9 or 10. At oestrus, AI was carried out twice, 12 h apart, with a total of 4 straws of X-sorted semen per cow. In the OPU group, cows (n = 7) were subjected to OPU without any pretreatment, collected immature oocytes were in vivo matured for 20 to 22 h, followed by IVF using X-sorted sperm for 6 h; then, presumptive zygotes were in vitro cultured (IVC) for 9 days. In the in vivo-matured oocyte group (matured group), a CIDR was inserted (Day 0) in cows (n = 4), all follicles larger than 8 mm were removed on Day 5. Administration of FSH, prostaglandin F2α, and GnRH, as well as withdrawal of CIDR, were performed as in the ER group. In vivo-matured oocytes were collected from follicles larger than 5 mm by OPU at 25 to 26 h following GnRH injection; collected oocytes with expanded cumulus cells were fertilized with X-sorted sperm 30 h after GnRH. After 6 h of IVF, presumptive zygotes were transferred to in vitro culture, as in the OPU group. Data were compared among 3 groups; the ER group was analysed for number of CL, collected embryos, and normal embryos, against the number of aspirated follicles, collected oocytes used for IVF, and formed blastocysts in the OPU and matured groups, respectively, by Tukey-kramer test after ANOVA. There were no differences between the number of CL in the ER group and the number of follicles in the OPU and matured groups (16.4 ± 5.3 v. 31.6 ± 22.7 v. 18.5 ± 4.7, mean ± s.d., respectively). Also the number of collected embryos in the ER group and number of oocytes for IVF in the OPU and matured groups (12.8 ± 7.6 v. 14.9 ± 11.8 v. 17.8 ± 7.7, respectively) was similar. However, the number of blastocysts in the matured group (13.0 ± 5.9; P < 0.01) was higher than that in the OPU group (3.0 ± 2.2) and in the ER group (2.8 ± 3.7). For female embryo production in Brown Swiss cows using X-sorted sperm, the system of IVF with in vivo matured oocytes obtained by OPU is more effective than ER or OPU without pretreatment.

scholarly journals Ex vivo Retrieval of Mature Oocytes for Fertility Preservation in a Patient with Bilateral Borderline Ovarian Tumor

2021 ◽  
Author(s):  
Bruno Ramalho de Carvalho ◽  
Geórgia Fontes Cintra ◽  
Taise Moura Franceschi ◽  
Íris de Oliveira Cabral ◽  
Leandro Santos de Araújo Resende ◽  
...  
Keyword(s):  
Fertility Preservation ◽  
Ovarian Stimulation ◽  
Ovarian Tumor ◽  
Ex Vivo ◽  
Oocyte Retrieval ◽  
Ovarian Response ◽  
Malignant Cell ◽  
Borderline Ovarian Tumor ◽  
Increased Risk

AbstractWe report a case of ultrasound-guided ex vivo oocyte retrieval for fertility preservation in a woman with bilateral borderline ovarian tumor, for whom conventional transvaginal oocyte retrieval was deemed unsafe because of the increased risk of malignant cell spillage. Ovarian stimulation with gonadotropins was performed. Surgery was scheduled according to the ovarian response to exogenous gonadotropic stimulation; oophorectomized specimens were obtained by laparoscopy, and oocyte retrieval was performed ∼ 37 hours after the ovulatory trigger. The sum of 20 ovarian follicles were aspirated, and 16 oocytes were obtained. We performed vitrification of 12 metaphase II oocytes and 3 oocytes matured in vitro. Our result emphasizes the viability of ex vivo mature oocyte retrieval after controlled ovarian stimulation for those with high risk of malignant dissemination by conventional approach.

scholarly journals L-OPU in Goat and Sheep—Different Variants of the Oocyte Recovery Method

Animals ◽  
2020 ◽  
Vol 10 (4) ◽  
pp. 658
Author(s):  
Jarosław Wieczorek ◽  
Jurij Koseniuk ◽  
Maria Skrzyszowska ◽  
Mirosław Cegła
Keyword(s):  
In Vitro Maturation ◽  
Hormonal Stimulation ◽  
Recovery Method ◽  
Vitro Fertilization ◽  
Minimal Invasiveness ◽  
Animal Reproduction ◽  
Oocyte Recovery ◽  
Maturation In Vitro ◽  
Invasive Techniques

The laparoscopic method of recovering oocytes in goats and sheep is one of the minimally invasive methods used in the biotechnology of animal reproduction. It allows for good quality oocytes that are suitable for in vitro maturation and fertilization to be recovered. The limitation of using the laparoscopic ovum pick-up (L-OPU) method in goat and sheep is its changing effectiveness and the lack of repeatability of results, as well as the varying effectiveness of different variants of the method. Therefore, it is necessary to develop effective non-invasive techniques allowing for multiple good quality oocyte recovery that would be suitable for in vitro maturation and fertilization. In this study, four different L-OPU variants were described in goats and sheep. Various techniques of recovering oocytes were discussed, including the techniques of conducting the operation, various tools for recovering oocytes, and different plans of hormonal stimulation. Recovery rates were 35% (Variant I), 57% (Variant II), 72% (Variant III), and 67% (Variant IV). After evaluation, 94% (both Variant I and II), 93% (Variant III), and 84% (Variant IV) of the oocytes were qualified for in vitro maturation. The results of the study show that the proposed technique of laparoscopic recovery of oocytes allows a sufficient number of ovarian cells suitable for in vitro culture to be obtained and as a consequence it makes them useful in in vitro maturation/in vitro fertilization (IVM/IVF) programs or cloning. The method allows for a fast and effective conduct of the operation in a living donor with minimal invasiveness while preserving the excellent condition of animals.

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